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目的:探讨人生长激素(hGH)基因修饰的鼠骨骼肌成肌细胞移植对心肌梗死大鼠血流动力学和血管新生的影响。方法:结扎大鼠左冠状动脉前降支制作大鼠心力衰竭模型。2周后将冠脉结扎后存活的30只大鼠随机分为3组,hGH基因修饰的成肌细胞移植组(hGH组)、绿色荧光蛋白(GFP)基因修饰的成肌细胞移植组(GFP组)、注射等体积培养液的对照组。治疗4周后,血流动力学检查各组心功能指标;心肌组织进行HE染色检测心肌梗死面积,Ⅷ因子免疫组织化学检测血管新生。RT-PCR检测bax和bcl-2 mRNA的表达。Western blot-ting检测各组心肌hGH、血管内皮生长因子(VEGF)、caspase-3蛋白表达情况。结果:(1)与对照组和GFP组相比:hGH组血流动力学指标明显改善,心肌梗死面积缩小。(2)心肌组织Ⅷ因子免疫组织化学检查结果显示:hGH组血管密度显著高于GFP组和对照组。(3)RT-PCR检测结果显示hGH组bax mRNA水平显著低于GFP组和对照组,bcl-2 mRNA水平显著高于GFP组和对照组。(4)Western blotting检测结果显示hGH组大鼠心肌组织有hGH蛋白表达,其余两组心肌组织无hGH蛋白表达。与其它两组相比,hGH组caspase-3蛋白表达降低,VEGF蛋白表达增加。结论:hGH基因修饰的成肌细胞移植可以抑制细胞凋亡,与单独成肌细胞治疗相比可以诱导更大的血管化,更好地缩小心肌梗死面积,并改善心肌梗死大鼠血流动力学。成肌细胞治疗联合hGH基因治疗为心肌梗死后心力衰竭的治疗提供了一个新的途径。
Objective: To investigate the effects of hGH gene-modified mouse skeletal myoblast transplantation on hemodynamics and angiogenesis in rats with myocardial infarction. Methods: Rat model of heart failure was established by ligation of left anterior descending coronary artery in rats. After 2 weeks, 30 rats surviving coronary artery ligation were randomly divided into 3 groups: hGH gene modified myoblast transplantation group (hGH group), green fluorescent protein (GFP) gene modified myoblast transplantation group (GFP Group), injection of equal volume of culture medium control group. After 4 weeks of treatment, hemodynamic examination of each group cardiac function indicators; myocardial tissue myocardial infarction area HE staining, Ⅷ factor immunohistochemical detection of angiogenesis. RT-PCR detection of bax and bcl-2 mRNA expression. Western blot-ting was used to detect the expression of hGH, VEGF and caspase-3 in each group. Results: (1) Compared with the control group and the GFP group, the hemodynamic indexes of hGH group were significantly improved and the area of myocardial infarction was reduced. (2) The results of immunohistochemistry of factor Ⅷ of myocardial tissue showed that the vascular density of hGH group was significantly higher than that of GFP group and control group. (3) The results of RT-PCR showed that the level of bax mRNA in hGH group was significantly lower than that in GFP group and control group, and the level of bcl-2 mRNA was significantly higher than that in GFP group and control group. (4) The results of Western blotting showed that hGH protein was expressed in the myocardium of hGH group, but no hGH protein in the other two groups. Compared with the other two groups, hGH group caspase-3 protein expression decreased, VEGF protein expression increased. CONCLUSIONS: hGH gene-modified myoblast transplantation can inhibit apoptosis and induce more vascularization than single myoblasts alone, thus reducing myocardial infarct size and improving hemodynamics in myocardial infarction rats . Myoblast therapy combined with hGH gene therapy provides a new avenue for the treatment of heart failure after myocardial infarction.